在皮升级液滴中创建逐步浓度梯度，用于基质金属蛋白酶 II 和 IX 的平行反应。
Creation of stepwise concentration gradient in picoliter droplets for parallel reactions of matrix metalloproteinase II and IX.
机构信息
Materials Research and Education Center, Department of Mechanical Engineering, Auburn University, Alabama 36849, United States.
出版信息
Anal Chem. 2011 May 1;83(9):3358-64. doi: 10.1021/ac103217p. Epub 2011 Apr 13.
Abstract
We present a new methodology for generating a stepwise concentration gradient in a series of microdroplets by using monolithic micro valves that act as "faucets" in micrometer-scale. A distinct concentration gradient of a substrate was generated for the determination of the kinetic parameters of two different enzymes using only 10 picoliter-scale droplets. With a single experiment on a chip, we obtained K(M) and k(cat) values of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9), and compared the catalytic competence of the two enzymes. The present system and method are highly suitable for applications where the reagents or samples are limited and precious.
摘要
我们提出了一种新的方法，通过使用整体式微阀在一系列微滴中产生逐步浓度梯度，微阀在微米尺度上充当“水龙头”。通过仅使用 10 皮升规模的液滴，为两种不同酶的动力学参数的测定生成了明显的浓度梯度。通过在芯片上进行单个实验，我们获得了基质金属蛋白酶 2 (MMP-2)和基质金属蛋白酶 9 (MMP-9)的 K(M)和 k(cat)值，并比较了两种酶的催化能力。本系统和方法非常适用于试剂或样品有限且珍贵的应用。
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